A Model of Piezo1-Based Regulation of Red Blood Cell Volume

A red blood cell (RBC) performs its function of adequately carrying respiratory gases in blood by its volume being ～60% of that of a sphere with the same membrane area. For this purpose, human and most other vertebrate RBCs regulate their content of potassium (K⁺) and sodium (Na⁺) ions. The focus considered here is on K⁺ efflux through calcium-ion (Ca²⁺)-activated Gárdos channels. These channels open under conditions that allow Ca²⁺ to enter RBCs through Piezo1 mechanosensitive cation-permeable channels. It is postulated that the fraction of open Piezo1 channels depends on the RBC shape as a result of the curvature-dependent Piezo1-bilayer membrane interaction. The consequences of this postulate are studied by introducing a simple model of RBC osmotic behavior supplemented by the dependence of RBC membrane K⁺ permeability on the reduced volume (i.e., the ratio of cell volume to its maximal possible volume) of RBC discoid shapes. It is assumed that because of its intrinsic curvature and strong interaction with the surrounding membrane, Piezo1 tends to concentrate in the dimple regions of these shapes, and the fraction of open Piezo1 channels depends on the membrane curvature in that region. It is shown that the properties of the described model can provide the basis for the formation of the negative feedback loop that interrelates cell volume and its content of potassium ions. The model predicts the relation, valid for each cell in an RBC population, between RBC volume and membrane area, thus explaining the large value of the measured membrane area versus the volume correlation coefficient. The mechanism proposed here for RBC volume regulation is in accord with the loss of this correlation in RBCs of Piezo1 knockout mice.     

Reconstruction of tricarboxylic acid cycle in Corynebacterium glutamicum with a genome-scale metabolic network model for trans-4-hydroxyproline production

trans-4-Hydroxy- l -proline (Hyp) is an abundant component of mammalian collagen and functions as a chiral synthon for the syntheses of anti-inflammatory drugs in the pharmaceutical industry. Proline 4-hydroxylase (P4H) can catalyze the conversion of l -proline to Hyp; however, it is still challenging for the fermentative production of Hyp from glucose using P4H due to the low yield and productivity. Here, we report the metabolic engineering of Corynebacterium glutamicum for the fermentative production of Hyp by reconstructing tricarboxylic acid (TCA) cycle together with heterologously expressing the p4h gene from Dactylosporangium sp. strain RH1. In silico model-based simulation showed that α-ketoglutarate was redirected from the TCA cycle toward Hyp synthetic pathway driven by P4H when the carbon flux from succinyl-CoA to succinate descended to zero. The interruption of the TCA cycle by the deletion of sucCD-encoding the succinyl-CoA synthetase (SUCOAS) led to a 60% increase in Hyp production and had no obvious impact on the growth rate. Fine-tuning of plasmid-borne ProB* and P4H abundances led to a significant increase in the yield of Hyp on glucose. The final engineered Hyp-7 strain produced up to 21.72 g/L Hyp with a yield of 0.27 mol/mol (Hyp/glucose) and a volumetric productivity of 0.36 g·L ⁻¹·hr ⁻¹ in the shake flask fermentation. To our knowledge, this is the highest yield and productivity achieved by microbial fermentation in a glucose-minimal medium for Hyp production. This strategy provides new insights into engineering C. glutamicum by flux coupling for the fermentative production of Hyp and related products.     

Circular RNA circMTO1 suppresses bladder cancer metastasis by sponging miR-221 and inhibiting epithelial-to-mesenchymal transition

Bladder cancer remains a leading cause of cancer-related death because of its distant metastasis and high recurrence rates. Deregulation of circular RNAs (circRNAs) can act either as tumor suppressors or oncogenes to control cell proliferation, migration, and metastasis. The role of circMTO1 in bladder cancer remain unknown. In this study, we investigated whether circMTO1 could use as a biomarker and therapeutic target for bladder cancer treatment. We first demonstrated that circMTO1 was an important circRNA frequently downregulated in bladder cancer tissue, and lower circMTO1 levels were positively correlated with bladder cancer patients' metastasis and poorer survival. Ectopic expression of circMTO1 in bladder cancer cells inhibited cell's epithelial-to-mesenchymal transition (EMT) and metastasis. In addition, we also revealed that circMTO1 was able to sponge miR-221 and overexpression of circMTO1 negatively regulated the E-cadherin/N-cadherin pathway to inhibit bladder cancer cells' EMT by competing for miR-221. In conclusion, our findings provide comprehensive evidences that circMTO1 is a prognostic biomarker in bladder cancer and suggest that circMTO1 may function as a novel therapeutic target in human bladder cancer.     

Interleukin-2 induces extracellular matrix synthesis and TGF-β2 expression in retinal pigment epithelial cells

Macular fibrosis is a vital obstacle of vision acuity improvement of age-related macular degeneration patients. This study was to investigate the effects of interleukin 2 (IL-2) on epithelial-mesenchymal transition (EMT), extracellular matrix (ECM) synthesis and transforming growth factor β2 (TGF-β2) expression in retinal pigment epithelial (RPE) cells. 10 μg/L IL-2 was used to induce fibrosis in RPE cells for various times. Western blot was used to detect the EMT marker α-smooth muscle actin (α-SMA), ECM markers fibronectin (Fn) and type 1 collagen (COL-1), TGF-β2, and the activation of the JAK/STAT3 and NF-κB signaling pathway. Furthermore, JAK/STAT3 and NF-κB signaling pathways were specifically blocked by WP1066 or BAY11-7082, respectively, and the expression of α-SMA, COL-1, Fn and TGF-β2 protein were detected. Wound healing and Transwell assays were used to measure cell migration ability of IL-2 with or without WP1066 or BAY11-7082. After induction of IL-2, the expressions of Fn, COL-1, TGF-β2 protein were significantly increased, and this effect was correlated with IL-2 treatment duration, while α-SMA protein expression did not change significantly. Both WP1066 and BAY11-7082 could effectively downregulate the expression of Fn, COL-1 and TGF-β2 induced by IL-2. What's more, both NF-κB and JAK/STAT3 inhibitors could suppress the activation of the other signaling pathway. Additionally, JAK/STAT3 inhibitor WP1066 and NF-κB inhibitor BAY 11-7082 could obviously decrease RPE cells migration capability induced by IL-2. IL-2 promotes cell migration, ECM synthesis and TGF-β2 expression in RPE cells via JAK/STAT3 and NF-κB signaling pathways, which may play an important role in proliferative vitreoretinopathy.     

Regulation by Hsp27/P53 in testis development and sperm apoptosis of male cattle (cattle-yak and yak)

Heat shock protein 27 (Hsp27)/protein 53 (P53) plays an important role in testis development and spermatozoa regulation, but the relationship between Hsp27/P53 and infertility in cattle is unclear. Here, we focus on male cattle-yak and yak to investigate the expression and localization of Hsp27/P53 in testis tissues and to explore the influence of Hsp27/P53 on infertility. In our study, a total of 54 cattle (24 cattle-yak and 30 yak) were examined. The Hsp27 and P53 messenger RNA (mRNA) of cattle-yak were cloned, and amino acid variations in Hsp27 and P53 were found; the variations led to differences in the protein spatial structure compared with yak. We used real-time quantitative polymerase chain reaction and western blot to investigate whether the expression of Hsp27/P53 mRNA and protein was different in cattle-yak and yak. We found that the expression levels of Hsp27/P53 mRNA and protein were different in the testis developmental stages and the highest expression was observed in testicles during adulthood. Moreover, the Hsp27 expression was significantly higher in yak, whereas P53 expression was higher in cattle-yak (p < 0.01). On this basis, we detected the location of Hsp27/P53 in the testis by immunohistochemistry and immunofluorescence. The results demonstrated that Hsp27 was located in spermatogenic cells at different developmental stages and mesenchymal cells of the yak testicles. However, P53 was located in the primary spermatocyte and interstitial cells of the cattle-yak testicles. In summary, our study proved that the expression of Hsp27/P53 differed across the testis developmental stages and the expression of P53 was higher in the testis of cattle-yak, which suggested that the infertility of cattle-yak may be caused by the upregulation of P53.     

Benzo(a)pyrene inhibits endometrial cell apoptosis in early pregnant mice via the WNT5A pathway

Benzo(a)pyrene (BaP) is an endocrine-disrupting pollutant present in various aspects of daily life, and studies have demonstrated that BaP exerts reproductive toxicity. We previously showed that BaP damages endometrial morphology and decreases the number of implantation sites in early pregnant mice, but the mechanisms underlying these effects remain unclear. The endometrial function is crucial for implantation, which is associated with endometrial cell apoptosis. In this study, we focused on the effect of BaP on endometrial cell apoptosis and the role of WNT signaling during this process. Pregnant mice were gavaged with corn oil (control group) or 0.2 mg·kg⁻¹·day ⁻¹ BaP (treatment group) from Days 1 to 6 of pregnancy. BaP impaired endometrial function by decreasing the expression of HOXA10 and BMP2, two markers of receptivity and decidualization. WNT5A and β-catenin were activated in the BaP group. BaP affected the expression of apoptosis-related proteins and inhibited the apoptosis of endometrial stromal cells. In vitro, human endometrial stromal cells (HESCs) were treated with different concentrations of BaP (dimethyl sulfoxide (DMSO); 5, 10 µM). WNT5A and β-catenin were also upregulated in the BaP treatment group. HESC apoptosis was restrained by BaP. Inhibiting WNT5A by SFRP5 partially restored the effect of BaP on apoptosis. In summary, these results suggested that BaP exposure during early pregnancy activates WNT5A/β-catenin signaling pathway, which inhibits the endometrial cell apoptosis and potentially destroys endometrial function.     

Alleviation of the detrimental effect of water deficit on wheat (Triticum aestivum L.) growth by an indole acetic acid-producing endophytic fungus

Plant and Soil - Endophytic fungi colonization is an eco-friendly strategy to respond to environmental stresses and confer tolerance to the host plant. Here, the responses of wheat plant inoculated...     

Paecilomyces variotii extracts (ZNC) enhance plant immunity and promote plant growth

Plant and Soil - There are growing concerns regarding the restoration of karst rocky desertification (KRD) areas. However, the soil conditions and its residing microorganisms, which are essential...     

Megabase Length Hypermutation Accompanies Human Structural Variation at 17p11.2

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